【摘要】 目的观察大黄素对脂多糖(LPS)诱导的角膜基质细胞表达细胞因子的影响。方法原代培养角膜基质细胞,选取第4代细胞进行实验。根据在LPS刺激角膜基质细胞前是否先用40μmol/L大黄素培养细胞30 min,将细胞分为大黄素+LPS组和LPS组,分别在10μg/LLPS刺激角膜基质细胞前以及刺激后1、2、4、8 h收集细胞;或先用0、5、10、20和40μmol/L的大黄素培养细胞30 min后,再用10μg/L LPS刺激8 h。Western blot检测角膜基质细胞κB抑制因子-α(IκB-α)蛋白的表达;逆转录聚合酶链反应(RT-PCR)检测角膜基质细胞白细胞介素(IL)-6和IL-8的mR-NA表达。结果LPS刺激角膜基质细胞后1、2、4、8 h,细胞表达IκB-α蛋白均较刺激前明显下降(P<0.01);大黄素对LPS引起的角膜基质细胞IκB-α蛋白的降解有不同程度的抑制作用,且该作用呈一定的浓度依赖性(P<0.05)。LPS刺激引起角膜基质细胞IL-6和IL-8 mRNA表达升高,刺激后各时间点IL-6和IL-8 mRNA表达均较LPS刺激前明显升高,呈一定时间依赖性(P<0.01);大黄素对LPS诱导的角膜基质细胞IL-6和IL-8 mRNA表达有明显抑制作用,且该作用呈一定浓度依赖性(P<0.05)。结论大黄素能抑制体外培养的角膜基质细胞表达IL-6和IL-8,可能与其抑制IκB-α降解、促进NF-kB活化有关。更多还原

【Abstract】 Objective To investigate the effects of emodin on expression of cytokines induced by lipopolysaccharide(LPS) in cultured human corneal fibroblasts in vitr o.Methods Primary human corneal fibroblasts of passages 4 were used in this research.Cells were treated with 10 μg/L LPS for 1,2,4,or 8 hours,which were pretreated with or without emodin for 30 minutes before LPS challenge.The degeneration of inhibitor of κB-α(IκB-α) and the effect of emodin on it were analyzed by Western blot analysis with a specific antibody.The cellular abundance of the mRNA of interleukin(IL)-6 and IL-8 from corneal fibroblasts under different conditions was determined by reverse transcriptase polymerase chain reaction(RT-PCR).Results Compared with cells without LPS treatment,IκB-α level significantly decreased in every time point after LPS challenge(P <0.01).Emodin inhibited the LPS-induced degeneration of IκB-α by corneal fibroblasts in a dose-dependent manner(P <0.05).Compared with cells without LPS treatment,the expressions of IL-6 and IL-8 mRNA significantly increased in every time point after LPS challenge(P <0.01).At the same time,the expressions of the mRNA of IL-6 and IL-8 induced by LPS in corneal fibroblasts were also inhibited by emodin in a dose-dependent manner(P <0.05).Conclusion Emodin can inhibit the expressions of IL-6 and IL-8 mRNA induced by LPS in corneal fibroblasts,which maybe via inhibiting the degeneration of IκB-α and suppressing the activation of nuclear factor-κB.更多还原